Antibody Profiling by Proteome Microarray with Multiplex Isotype Detection Reveals Overlap between Human and Aotus nancymaae Controlled Malaria Infections

Abstract

The development of vaccines against malaria and serodiagnostic tests for detecting recent exposure requires tools for antigen discovery and suitable animal models. The protein microarray is a high-throughput, sample sparing technique, with applications in infectious disease research, clinical diagnostics, epidemiology, and vaccine development. We recently demonstrated Qdot-based indirect immunofluorescence together with portable optical imager ArrayCAM using single isotype detection could replicate data using the conventional laser confocal scanner system. We developed a multiplexing protocol for simultaneous detection of IgG, IgA, and IgM and compared samples from a controlled human malaria infection model with those from controlled malaria infections of Aotus nancymaae, a widely used non-human primate model of human malaria. IgG profiles showed the highest concordance in number of reactive antigens; thus, of the 139 antigens recognized by human IgG antibody, 111 were also recognized by Aotus monkeys. Interestingly, IgA profiles were largely non-overlapping. Finally, on the path toward wider deployment of the portable platform, we show excellent correlations between array data obtained in five independent laboratories around the United States using the multiplexing protocol (R : 0.60-0.92). This study supports the use of this platform for wider deployment, particularly in endemic areas where such a tool will have the greatest impact on global human health.

Administration of Multivalent Influenza Virus Recombinant Hemagglutinin Vaccine in Combination-Adjuvant Elicits Broad Reactivity Beyond the Vaccine Components

“…HA protein microarrays were fabricated as previously described (52,53). Briefly, lyophilized influenza proteins (Sino Biological Inc., Beijing, China) were reconstituted to a concentration of 0.1 mg/ml in phosphate-buffered saline (PBS) with 0.001% Tween 20 (T-PBS) and printed onto nitrocellulose-coated glass Oncyte ® Avid slides (Grace Bio-Labs, Inc., Bend, OR) using an Omni Grid 100 microarray printer (Genomic Solutions).…”

Distinct Antibody Signatures Associated with Different Malaria Transmission Intensities in Zambia and Zimbabwe

“…To this end, we recently validated a portable digital complementary metal oxide semiconductor (CMOS) array imaging system as a low-cost and robust alternative that could easily be deployed in laboratory settings where the purchase and maintenance of a more complex machine would be cost prohibitive or impractical (44). We also reported that the probing protocol was readily translated to other labs, as shown by excellent correlations between the data generated by five independent laboratories (45). With the availability of training and commercial sources of arrays, the microarray technology can be disseminated more widely, particularly in areas of endemicity, where it will have the greatest impact on public health.…”

Development of a Multiplex Glycan Microarray Assay and Comparative Analysis of Human Serum Anti-Glycan IgA, IgG, and IgM Repertoires

“…For example, Taghavian et al state that signal loss was not a problem in their assay based on good Pearson correlation constants; however, the data show a variety of multiplex IgA signals that are lower than expected and the slopes of the lines were lower than 1.0 ( y = 0.84), indicating an overall trend of lower than expected signals for the multiplex assay. 19…”